背景:由于相对丰度数据的半定性性质,使用下一代DNA测序的转化微生物组研究具有挑战性。在这项为期12周的临床试验中应用了一种新的定量分析方法,以了解机械与刷牙的化疗作用,使用牙线,并对牙龈上的牙菌斑微生物组进行口腔冲洗。还使用vPCR对牙龈上菌斑进行了计数,以进行验证,并对牙龈下菌斑进行了计数,以评估牙龈边缘以下的干预效果。
方法:患牙龈炎的受试者纳入单一中心,检查官盲,虚拟监督,平行组对照临床试验。患有牙龈炎的受试者被随机分为仅刷牙(B);刷牙和使用牙线(BF);用Listerine®CoolMint®防腐剂(BA)刷牙和冲洗;用Listerine®CoolMint®Zero(BZ)刷牙和冲洗;或刷牙,使用牙线,并用Listerine®CoolMint®Zero(BFZ)冲洗。所有受试者每天用单氟磷酸钠牙膏和软毛牙刷刷牙两次,持续1分钟。牙线的受试者每天使用一次无味的上蜡牙线。分配给漱口水的受试者每天冲洗两次。在基线访视以及干预4周和12周后收集斑块标本。通过在DNA提取之前将参考量的DNA对照添加到噬斑样品中来实现细菌细胞数量定量。其次是浅鸟枪宏基因组测序。
结果:286名受试者完成了试验。牙龈上斑块的宏基因组数据显示Shannon-Weaver多样性显着降低,物种丰富度,以及总细菌和分类细菌丰度(共生,牙龈炎,和恶臭)在BA的4周和12周后,BZ,和BFZ组与B组相比,B组和BF组之间无显著差异。牙龈上菌斑vPCR进一步验证了这些结果,牙龈下菌斑vPCR仅对BFZ干预具有显着的功效。
结论:本出版物报道了微生物组分析的定量方法在临床试验中的成功应用,证明与机械方法相比,精油漱口水在控制牙菌斑方面具有持续和优异的功效。该试验中的定量微生物数据还加强了EO漱口剂对牙菌斑微生物生态的安全性和作用机制,并强调了提高EO漱口剂作为口腔卫生方案不可或缺的一部分的重要性。
背景:该试验于2022年10月31日在ClinicalTrials.gov上注册。注册号是NCT05600231。
BACKGROUND: Translational microbiome research using next-generation DNA sequencing is challenging due to the semi-qualitative nature of relative abundance data. A novel method for quantitative analysis was applied in this 12-week clinical trial to understand the mechanical vs. chemotherapeutic actions of brushing, flossing, and mouthrinsing against the supragingival dental plaque microbiome. Enumeration of viable bacteria using vPCR was also applied on supragingival plaque for validation and on subgingival plaque to evaluate interventional effects below the gingival margin.
METHODS: Subjects with gingivitis were enrolled in a single center, examiner-blind, virtually supervised, parallel group controlled clinical trial. Subjects with gingivitis were randomized into brushing only (B); brushing and flossing (BF); brushing and rinsing with Listerine® Cool Mint® Antiseptic (BA); brushing and rinsing with Listerine® Cool Mint® Zero (BZ); or brushing, flossing, and rinsing with Listerine® Cool Mint® Zero (BFZ). All subjects brushed twice daily for 1 min with a sodium monofluorophosphate toothpaste and a soft-bristled toothbrush. Subjects who flossed used unflavored waxed dental floss once daily. Subjects assigned to mouthrinses rinsed twice daily. Plaque specimens were collected at the baseline visit and after 4 and 12 weeks of intervention. Bacterial cell number quantification was achieved by adding reference amounts of DNA controls to plaque samples prior to DNA extraction, followed by shallow shotgun metagenome sequencing.
RESULTS: 286 subjects completed the trial. The metagenomic data for supragingival plaque showed significant reductions in Shannon-Weaver diversity, species richness, and total and categorical bacterial abundances (commensal, gingivitis, and malodor) after 4 and 12 weeks for the BA, BZ, and BFZ groups compared to the B group, while no significant differences were observed between the B and BF groups. Supragingival plaque vPCR further validated these results, and subgingival plaque vPCR demonstrated significant efficacy for the BFZ intervention only.
CONCLUSIONS: This publication reports on a successful application of a quantitative method of microbiome analysis in a clinical trial demonstrating the sustained and superior efficacy of essential oil mouthrinses at controlling dental plaque compared to mechanical methods. The quantitative microbiological data in this trial also reinforce the safety and mechanism of action of EO mouthrinses against plaque microbial ecology and highlights the importance of elevating EO mouthrinsing as an integral part of an oral hygiene regimen.
BACKGROUND: The trial was registered on ClinicalTrials.gov on 31/10/2022. The registration number is NCT05600231.